In the past several years, the advent of recombinant DNA technology was seen as a revolution of biotechnology. Recombinant DNA, also known as rDNA, is an artificial type of DNA which formed by combining two or more sequences and strands of DNA that would not occur together usually. rDNA is sometimes called genetic engineering. The applications of recombinant DNA technology are widely used and increasing recently for various kinds of purposes for example, expand the potential of microorganisms, to produce natural proteins manufacture of vaccines or enzymes. Recombinant DNA is made by three common ways which included the transformation, phage introduction and non-bacterial transformation. Transformation is the step of inserting the interest DNA into a vector and then cut that piece of DNA with a restricted enzyme and then insert the DNA into the vector with DNA Ligase. After that the vector is inserted to a host cell like E. coli. The phage introduction is the process of transfection, which can be said as an equivalent of transformation but instead of using bacteria, a phage is used. This process uses lambda or other phages to produce phage plaques which contain recombinant. In non-bacterial transformation, a process which very is similar to transformation never use bacterial for the host. In microinjection, the DNA is injected directly into the nucleus of the cell being transformed while in biolistics, the host cells are bombarded with high velocity of microprojectiles. This technique was discovered by Peter Lobban and A. Dale Kaiser in Stanford University.
An overview of rDNA procedures
First of all, the vector should be isolated from an organism such as bacterium. Then the interest gene is inserted into that particular vector. The vector that chosen must be a self-replicating type of vector. As the interest gene was inserted into the vector, its taken up by a cell for example a bacterium. The cell is become transformed bacterium and the recombinant vector is grown to form many copies of clone of that vector. After finished cloning, the cells may isolated by the researchers to use for many purposes. One of the process that might undergo by the cloned cells is inserted into an organism to express in it and produce protein as the product. Moreover, a succeed of production of a rDNA needs some tools or techniques. Restriction enzymes are one of the tools. It is a special class of DNA-cutting enzyme which can recognizes and cuts or even digest one particular sequence of interest DNA. ( For example BamHI, EcorRI, HaeIII and HindIII ). As we know DNA is appear in double stranded and these enzymes cut both strands of DNA and produce blunt ends and staggered cuts which is not directly opposite each other. These ends can used to join two pieces of DNA and these sticky ends stretches single DNA by complementary base pairing.
Besides, vectors are another tools which used in rDNA technology. Vectors is seen as a vehicle for the replication of interest DNA sequences. It needs to be able for self replicating and to be a size for manipulated outside the cell along the rDNA process. Smaller vectors are easier to be manipulated which is not fragile when compare to the larger one. Shuttle vectors which are plasmids that exist in various species can be used to move cloned DNA among organisms. One of the techniques in rDNA is Polymerase Chain Reaction (PCR). This process is used to amplify the size of samples and it is used to increase the amount of DNA into large enough for research in a short period. Firstly, every strand of target or interest DNA will be the template for DNA synthesis. Four nucleotides, enzymes for catalyzing purposes and DNA polymerase is added to this DNA to form a new DNA. Primers (short pieces of nucleic acid) is added too to enhance the reaction start. The primers are complementary to the interest DNA and it will hybridize to make the DNA amplified. Now is the time for the polymerase to synthesis new strands which are complementary. Finally, the DNA is heated to convert the new DNA into single strands.
What Are the Benefits of Recombinant DNA Technology?
One breakthrough in recombinant DNA technology was the manufacture of biosynthetic "human" insulin, which was the first medicine made through recombinant DNA technology ever to be approved by the FDA. Insulin was the ideal candidate because it is a relatively simple protein and was therefore relatively easy to copy, as well as being extensively used to the extent that if researchers could prove that biosynthetic "human" insulin was safe and effective, the technology would be accepted as such, and would open opportunities for other products to be made in this fashion.
The specific gene sequence, or oligonucleuotide , that codes for insulin production in humans was introduced to a sample colony of E.coli (the bacteria found in the human intestine). Only about 1 out of 106 bacteria picks up the sequence. However, because the lifecycle is only about 30 minutes for E. coli, this limitation is not problematic, and in a 24-hour period, there may be billions of E. coli that are coded with the DNA sequences needed to induce insulin production. However, a sampling of initial reaction showed that Humulin was greeted more as a technological rather than a medical breakthrough, and that this sentiment was building even before the drug reached pharmacies. Ultimately, widespread consumer adoption of biosynthetic "human" insulin did not occur until the manufacturers removed highly-purified animal insulin from the market, thereby leaving consumers with no other alternative to synthetic varieties.
Recombinant cells are produced by inserting foreign genes into genetic code, or DNA. The process of recombination involves a vector, or gene carrier, that is inserted into a host cell. From this process, a variety of DNA technologies have been developed. The benefits of recombinant DNA include improvements in;
Cancer Research:
Through analyzing the genetic differences between normal cells and cancer cells, scientists are attempting to learn which genes are responsible for the uncontrolled growth of cancerous cells, as well as the ways in which these genes are activated or inactivated. According to geneticist Dr. James Frieson, it is possible to regulate a cell´s production of proteins by splicing portions of genetic code that affect that cell´s regulatory functions. If this method could be applied to cancerous cells through recombinant DNA technology, it may be able to halt uncontrolled cell growth.
Fertility:
Recombinant DNA technology is used to produce hormones for women with fertility issues. Recombinant human follicle stimulating hormone (r-hFSH), recombinant luteinizing hormone (r-hLH) and recombinant human chorionic gonadotropin (r-hCG) are all hormones that facilitate the proper functioning of ovulation and follicular maturation necessary for fertilization to become a success. As opposed to earlier methods of hormone production, recombinant DNA technology will bring about a higher efficacy, easier access and safer, less invasive infertility treatments.
Vaccines:
Recombinant DNA is used in vaccines that involve the direct injection of genetic material into the human body. This genetic material is in the form of a plasmid, or loop of DNA, from the foreign antigen that is the target of the vaccination. After it is injected through our muscle tissue, our cells take in the DNA and begin to produce the foreign proteins encoded in the plasmids. These proteins promote our bodies´ immune responses to the targeted antigen. DNA vaccinations could become less costly to produce, are potentially safer and are theoretically longer lasting than alternative forms of vaccinations.
Diabetes Treatment:
Recombinant DNA can be used to treat a variety of other diseases and conditions. The production of insulin through recombinant DNA technology has been especially effective for treating diabetes patients. Today, scientists are able to create human insulin that is identical to pancreatic insulin, thereby leading to the the safest and purest forms of insulin on the market.
Food Recombinant:
DNA has a role in food production for a number of plant and animal products. For crops, recombinant DNA has been used to create increased resistance to viruses/pests, more resilience in the face of harsh environmental conditions and added convenience for packaging and shipping. An example for use with animals is bovine somatotropin (bST), a hormone that can be bacterially inserted in dairy cows in order to increase milk production.
References:
1. Colowick, S. P.; Kapian, O. N. (1980). Methods in Enzymology - Volume 68; Recombinant DNA. Academic Press.
2. Tortora, Funke, Case. 2010. Biotechnology and Recombinant DNA. (10th edition). Pearson Education, Inc. Pg 247-270
An overview of rDNA procedures
First of all, the vector should be isolated from an organism such as bacterium. Then the interest gene is inserted into that particular vector. The vector that chosen must be a self-replicating type of vector. As the interest gene was inserted into the vector, its taken up by a cell for example a bacterium. The cell is become transformed bacterium and the recombinant vector is grown to form many copies of clone of that vector. After finished cloning, the cells may isolated by the researchers to use for many purposes. One of the process that might undergo by the cloned cells is inserted into an organism to express in it and produce protein as the product. Moreover, a succeed of production of a rDNA needs some tools or techniques. Restriction enzymes are one of the tools. It is a special class of DNA-cutting enzyme which can recognizes and cuts or even digest one particular sequence of interest DNA. ( For example BamHI, EcorRI, HaeIII and HindIII ). As we know DNA is appear in double stranded and these enzymes cut both strands of DNA and produce blunt ends and staggered cuts which is not directly opposite each other. These ends can used to join two pieces of DNA and these sticky ends stretches single DNA by complementary base pairing.
Besides, vectors are another tools which used in rDNA technology. Vectors is seen as a vehicle for the replication of interest DNA sequences. It needs to be able for self replicating and to be a size for manipulated outside the cell along the rDNA process. Smaller vectors are easier to be manipulated which is not fragile when compare to the larger one. Shuttle vectors which are plasmids that exist in various species can be used to move cloned DNA among organisms. One of the techniques in rDNA is Polymerase Chain Reaction (PCR). This process is used to amplify the size of samples and it is used to increase the amount of DNA into large enough for research in a short period. Firstly, every strand of target or interest DNA will be the template for DNA synthesis. Four nucleotides, enzymes for catalyzing purposes and DNA polymerase is added to this DNA to form a new DNA. Primers (short pieces of nucleic acid) is added too to enhance the reaction start. The primers are complementary to the interest DNA and it will hybridize to make the DNA amplified. Now is the time for the polymerase to synthesis new strands which are complementary. Finally, the DNA is heated to convert the new DNA into single strands.
What Are the Benefits of Recombinant DNA Technology?
One breakthrough in recombinant DNA technology was the manufacture of biosynthetic "human" insulin, which was the first medicine made through recombinant DNA technology ever to be approved by the FDA. Insulin was the ideal candidate because it is a relatively simple protein and was therefore relatively easy to copy, as well as being extensively used to the extent that if researchers could prove that biosynthetic "human" insulin was safe and effective, the technology would be accepted as such, and would open opportunities for other products to be made in this fashion.
The specific gene sequence, or oligonucleuotide , that codes for insulin production in humans was introduced to a sample colony of E.coli (the bacteria found in the human intestine). Only about 1 out of 106 bacteria picks up the sequence. However, because the lifecycle is only about 30 minutes for E. coli, this limitation is not problematic, and in a 24-hour period, there may be billions of E. coli that are coded with the DNA sequences needed to induce insulin production. However, a sampling of initial reaction showed that Humulin was greeted more as a technological rather than a medical breakthrough, and that this sentiment was building even before the drug reached pharmacies. Ultimately, widespread consumer adoption of biosynthetic "human" insulin did not occur until the manufacturers removed highly-purified animal insulin from the market, thereby leaving consumers with no other alternative to synthetic varieties.
Recombinant cells are produced by inserting foreign genes into genetic code, or DNA. The process of recombination involves a vector, or gene carrier, that is inserted into a host cell. From this process, a variety of DNA technologies have been developed. The benefits of recombinant DNA include improvements in;
Cancer Research:
Through analyzing the genetic differences between normal cells and cancer cells, scientists are attempting to learn which genes are responsible for the uncontrolled growth of cancerous cells, as well as the ways in which these genes are activated or inactivated. According to geneticist Dr. James Frieson, it is possible to regulate a cell´s production of proteins by splicing portions of genetic code that affect that cell´s regulatory functions. If this method could be applied to cancerous cells through recombinant DNA technology, it may be able to halt uncontrolled cell growth.
Fertility:
Recombinant DNA technology is used to produce hormones for women with fertility issues. Recombinant human follicle stimulating hormone (r-hFSH), recombinant luteinizing hormone (r-hLH) and recombinant human chorionic gonadotropin (r-hCG) are all hormones that facilitate the proper functioning of ovulation and follicular maturation necessary for fertilization to become a success. As opposed to earlier methods of hormone production, recombinant DNA technology will bring about a higher efficacy, easier access and safer, less invasive infertility treatments.
Vaccines:
Recombinant DNA is used in vaccines that involve the direct injection of genetic material into the human body. This genetic material is in the form of a plasmid, or loop of DNA, from the foreign antigen that is the target of the vaccination. After it is injected through our muscle tissue, our cells take in the DNA and begin to produce the foreign proteins encoded in the plasmids. These proteins promote our bodies´ immune responses to the targeted antigen. DNA vaccinations could become less costly to produce, are potentially safer and are theoretically longer lasting than alternative forms of vaccinations.
Diabetes Treatment:
Recombinant DNA can be used to treat a variety of other diseases and conditions. The production of insulin through recombinant DNA technology has been especially effective for treating diabetes patients. Today, scientists are able to create human insulin that is identical to pancreatic insulin, thereby leading to the the safest and purest forms of insulin on the market.
Food Recombinant:
DNA has a role in food production for a number of plant and animal products. For crops, recombinant DNA has been used to create increased resistance to viruses/pests, more resilience in the face of harsh environmental conditions and added convenience for packaging and shipping. An example for use with animals is bovine somatotropin (bST), a hormone that can be bacterially inserted in dairy cows in order to increase milk production.
References:
1. Colowick, S. P.; Kapian, O. N. (1980). Methods in Enzymology - Volume 68; Recombinant DNA. Academic Press.
2. Tortora, Funke, Case. 2010. Biotechnology and Recombinant DNA. (10th edition). Pearson Education, Inc. Pg 247-270
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